应激和分子伴侣对鱼藤酮诱导的SH-SY5Y细胞毒性的保护作用

目的:本文探讨应激和分子伴侣(stress and chaperone,STCH)对鱼藤酮的神经毒性之于SH-SY5Y细胞的保护作用及其可能的作用机制。方法:采用不同浓度鱼藤酮处理SH-SY5Y-pcDNA 3.1和SH-SY5Y-pcDNA3.1-STCH稳定细胞株,并观察细胞形态;采用MTT法和Western蛋白印迹法检测MAPK通路相关的信号分子和凋亡相关蛋白变化。结果:与未处理组相比,鱼藤酮均能引起细胞活力显著下降(P<0.05);STCH对鱼藤酮诱导的SH-SY5Y细胞毒性具有明显的保护作用(0.1μmol/L-5μmol/L),但10μmol/L浓度组与转染空载体组无显著差异。激活型caspase-3蛋白在鱼藤酮毒素处理后表达明显,p38磷酸化增强,过表达STCH在鱼藤酮处理后p38磷酸化受到明显抑制,裂解的caspase-3活性形式减少。结论:STCH对鱼藤酮诱导神经毒性的保护作用可能通过抑制p38磷酸化而实现。

Protective effect of STCH on rotenone induced neurotoxicity of human SH-SY5Y cells

Objective: We explored the protective effects and possible mechanism of stress and chaperone(STCH) on rotenone-induced neurotoxicity in SH-SY5Y.Methods: Human SH-SY5Y-pcDNA3.1 and SH-SY5Y-pcDNA3.1-STCH cells were treated with rotenone of different concentration,respectively;and the cell morphology was observed.The cell viability was evaluated by MTT assay and molecules related to MAPK signaling pathway and cell apoptosis were examined by Western Blot.Results: Our data showed that rotenone induced a decrease of cell viability in SH-SY5Y(P<0.05) compared to control.Compared to empty vector transfected group,STCH showed a protective role with concentrations ranging from 0.1 μmol/L to 5 μmol/L of rotenone(P<0.05) while there was no significant difference at 10 μmol/L of rotenone as demonstrated by SH-SY5Y-pcDNA3.1 and SH-SY5Y-pcDNA3.1-STCH cells groups.Compared to non-treatment group,increase of cleaved caspase-3 form and phosphorylation of p38 after treatment with rotenone in SH-SY5Y-pcDNA3.1 were detected respectively.While in SH-SY5Y-pcDNA3.1-STCH group(over-expression of STCH following rotenone treatment),activation of cleavage of caspase-3 form and phosphorylation of p38 were suppressed.Conclusion: These data indicate that the protective effects of STCH against rotenone-induced neurotoxicity may be mediated by suppressing activation of the p38 signaling-induced apoptosis.